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. 1990 Dec 1;111(6):2713–2723. doi: 10.1083/jcb.111.6.2713

Role of the extracellular matrix protein thrombospondin in the early development of the mouse embryo

PMCID: PMC2116366  PMID: 2277082

Abstract

The distribution of the extracellular matrix protein thrombospondin (TSP) in cleavage to egg cylinder staged mouse embryos and its role in trophoblast outgrowth from cultured blastocysts were examined. TSP was present within the cytoplasm of unfertilized eggs; in fertilized one- to four-cell embryos; by the eight-cell stage, TSP was also densely deposited at cell-cell borders. In the blastocyst, although TSP was present in all three cell types; trophectoderm, endoderm, and inner cell mass (ICM), it was enriched in the ICM and at the surface of trophectoderm cells. Hatched blastocysts grown on matrix-coated coverslips formed extensive trophoblast outgrowths on TSP, grew slightly less avidly on laminin, or on a 140-kD fragment of TSP containing its COOH terminus and putative cell binding domains. There was little outgrowth on the NH2 terminus heparin-binding domain. Addition of anti-TSP antibodies (but not GRGDS) to blastocysts growing on TSP strikingly inhibited outgrowth. Consistent with its early appearance and presence in trophoblast cells during implantation, TSP may play an important role in the early events involved in mammalian embryogenesis.

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Selected References

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  1. Armant D. R., Kaplan H. A., Lennarz W. J. Fibronectin and laminin promote in vitro attachment and outgrowth of mouse blastocysts. Dev Biol. 1986 Aug;116(2):519–523. doi: 10.1016/0012-1606(86)90152-1. [DOI] [PubMed] [Google Scholar]
  2. Armant D. R., Kaplan H. A., Mover H., Lennarz W. J. The effect of hexapeptides on attachment and outgrowth of mouse blastocysts cultured in vitro: evidence for the involvement of the cell recognition tripeptide Arg-Gly-Asp. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6751–6755. doi: 10.1073/pnas.83.18.6751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beers W. H. Follicular plasminogen and plasminogen activator and the effect of plasmin on ovarian follicle wall. Cell. 1975 Nov;6(3):379–386. doi: 10.1016/0092-8674(75)90187-7. [DOI] [PubMed] [Google Scholar]
  4. Damjanov I., Damjanov A., Damsky C. H. Developmentally regulated expression of the cell-cell adhesion glycoprotein cell-CAM 120/80 in peri-implantation mouse embryos and extraembryonic membranes. Dev Biol. 1986 Jul;116(1):194–202. doi: 10.1016/0012-1606(86)90056-4. [DOI] [PubMed] [Google Scholar]
  5. Damsky C. H., Richa J., Solter D., Knudsen K., Buck C. A. Identification and purification of a cell surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue. Cell. 1983 Sep;34(2):455–466. doi: 10.1016/0092-8674(83)90379-3. [DOI] [PubMed] [Google Scholar]
  6. Dixit V. M., Grant G. A., Santoro S. A., Frazier W. A. Isolation and characterization of a heparin-binding domain from the amino terminus of platelet thrombospondin. J Biol Chem. 1984 Aug 25;259(16):10100–10105. [PubMed] [Google Scholar]
  7. Dziadek M., Fujiwara S., Paulsson M., Timpl R. Immunological characterization of basement membrane types of heparan sulfate proteoglycan. EMBO J. 1985 Apr;4(4):905–912. doi: 10.1002/j.1460-2075.1985.tb03717.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dziadek M., Timpl R. Expression of nidogen and laminin in basement membranes during mouse embryogenesis and in teratocarcinoma cells. Dev Biol. 1985 Oct;111(2):372–382. doi: 10.1016/0012-1606(85)90491-9. [DOI] [PubMed] [Google Scholar]
  9. Farach M. C., Tang J. P., Decker G. L., Carson D. D. Heparin/heparan sulfate is involved in attachment and spreading of mouse embryos in vitro. Dev Biol. 1987 Oct;123(2):401–410. doi: 10.1016/0012-1606(87)90398-8. [DOI] [PubMed] [Google Scholar]
  10. Fisher S. J., Cui T. Y., Zhang L., Hartman L., Grahl K., Zhang G. Y., Tarpey J., Damsky C. H. Adhesive and degradative properties of human placental cytotrophoblast cells in vitro. J Cell Biol. 1989 Aug;109(2):891–902. doi: 10.1083/jcb.109.2.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huarte J., Belin D., Vassalli A., Strickland S., Vassalli J. D. Meiotic maturation of mouse oocytes triggers the translation and polyadenylation of dormant tissue-type plasminogen activator mRNA. Genes Dev. 1987 Dec;1(10):1201–1211. doi: 10.1101/gad.1.10.1201. [DOI] [PubMed] [Google Scholar]
  12. Huarte J., Belin D., Vassalli J. D. Plasminogen activator in mouse and rat oocytes: induction during meiotic maturation. Cell. 1985 Dec;43(2 Pt 1):551–558. doi: 10.1016/0092-8674(85)90184-9. [DOI] [PubMed] [Google Scholar]
  13. Kubo H., Spindle A., Pedersen R. A. Inhibition of mouse blastocyst attachment and outgrowth by protease inhibitors. J Exp Zool. 1981 Jun;216(3):445–451. doi: 10.1002/jez.1402160313. [DOI] [PubMed] [Google Scholar]
  14. Lahav J., Lawler J., Gimbrone M. A. Thrombospondin interactions with fibronectin and fibrinogen. Mutual inhibition in binding. Eur J Biochem. 1984 Nov 15;145(1):151–156. doi: 10.1111/j.1432-1033.1984.tb08534.x. [DOI] [PubMed] [Google Scholar]
  15. Lahav J., Schwartz M. A., Hynes R. O. Analysis of platelet adhesion with a radioactive chemical crosslinking reagent: interaction of thrombospondin with fibronectin and collagen. Cell. 1982 Nov;31(1):253–262. doi: 10.1016/0092-8674(82)90425-1. [DOI] [PubMed] [Google Scholar]
  16. Lawler J., Derick L. H., Connolly J. E., Chen J. H., Chao F. C. The structure of human platelet thrombospondin. J Biol Chem. 1985 Mar 25;260(6):3762–3772. [PubMed] [Google Scholar]
  17. Lawler J., Weinstein R., Hynes R. O. Cell attachment to thrombospondin: the role of ARG-GLY-ASP, calcium, and integrin receptors. J Cell Biol. 1988 Dec;107(6 Pt 1):2351–2361. doi: 10.1083/jcb.107.6.2351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leivo I., Vaheri A., Timpl R., Wartiovaara J. Appearance and distribution of collagens and laminin in the early mouse embryo. Dev Biol. 1980 Apr;76(1):100–114. doi: 10.1016/0012-1606(80)90365-6. [DOI] [PubMed] [Google Scholar]
  19. Liedholm P., Astedt B. Fibrinolytic activity of the rat ovum, appearance during tubal passage and disappearance at implantation. Int J Fertil. 1975;20(1):24–26. [PubMed] [Google Scholar]
  20. Liu Y. X., Cajander S. B., Ny T., Kristensen P., Hsueh A. J. Gonadotropin regulation of tissue-type and urokinase-type plasminogen activators in rat granulosa and theca-interstitial cells during the periovulatory period. Mol Cell Endocrinol. 1987 Dec;54(2-3):221–229. doi: 10.1016/0303-7207(87)90160-2. [DOI] [PubMed] [Google Scholar]
  21. Majack R. A., Cook S. C., Bornstein P. Control of smooth muscle cell growth by components of the extracellular matrix: autocrine role for thrombospondin. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9050–9054. doi: 10.1073/pnas.83.23.9050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McKeown-Longo P. J., Hanning R., Mosher D. F. Binding and degradation of platelet thrombospondin by cultured fibroblasts. J Cell Biol. 1984 Jan;98(1):22–28. doi: 10.1083/jcb.98.1.22. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mumby S. M., Raugi G. J., Bornstein P. Interactions of thrombospondin with extracellular matrix proteins: selective binding to type V collagen. J Cell Biol. 1984 Feb;98(2):646–652. doi: 10.1083/jcb.98.2.646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Murphy-Ullrich J. E., Hök M. Thrombospondin modulates focal adhesions in endothelial cells. J Cell Biol. 1989 Sep;109(3):1309–1319. doi: 10.1083/jcb.109.3.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Shea K. S., Dixit V. M. Unique distribution of the extracellular matrix component thrombospondin in the developing mouse embryo. J Cell Biol. 1988 Dec;107(6 Pt 2):2737–2748. doi: 10.1083/jcb.107.6.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Shea K. S., Rheinheimer J. S., Dixit V. M. Deposition and role of thrombospondin in the histogenesis of the cerebellar cortex. J Cell Biol. 1990 Apr;110(4):1275–1283. doi: 10.1083/jcb.110.4.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Prochownik E. V., O'Rourke K., Dixit V. M. Expression and analysis of COOH-terminal deletions of the human thrombospondin molecule. J Cell Biol. 1989 Aug;109(2):843–852. doi: 10.1083/jcb.109.2.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Queenan J. T., Jr, Kao L. C., Arboleda C. E., Ulloa-Aguirre A., Golos T. G., Cines D. B., Strauss J. F., 3rd Regulation of urokinase-type plasminogen activator production by cultured human cytotrophoblasts. J Biol Chem. 1987 Aug 15;262(23):10903–10906. [PubMed] [Google Scholar]
  29. Richa J., Damsky C. H., Buck C. A., Knowles B. B., Solter D. Cell surface glycoproteins mediate compaction, trophoblast attachment, and endoderm formation during early mouse development. Dev Biol. 1985 Apr;108(2):513–521. doi: 10.1016/0012-1606(85)90054-5. [DOI] [PubMed] [Google Scholar]
  30. Roberts D. D., Sherwood J. A., Ginsburg V. Platelet thrombospondin mediates attachment and spreading of human melanoma cells. J Cell Biol. 1987 Jan;104(1):131–139. doi: 10.1083/jcb.104.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  32. Saksela O., Rifkin D. B. Cell-associated plasminogen activation: regulation and physiological functions. Annu Rev Cell Biol. 1988;4:93–126. doi: 10.1146/annurev.cb.04.110188.000521. [DOI] [PubMed] [Google Scholar]
  33. Sappino A. P., Huarte J., Belin D., Vassalli J. D. Plasminogen activators in tissue remodeling and invasion: mRNA localization in mouse ovaries and implanting embryos. J Cell Biol. 1989 Nov;109(5):2471–2479. doi: 10.1083/jcb.109.5.2471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Silverstein R. L., Leung L. L., Harpel P. C., Nachman R. L. Complex formation of platelet thrombospondin with plasminogen. Modulation of activation by tissue activator. J Clin Invest. 1984 Nov;74(5):1625–1633. doi: 10.1172/JCI111578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Silverstein R. L., Nachman R. L., Leung L. L., Harpel P. C. Activation of immobilized plasminogen by tissue activator. Multimolecular complex formation. J Biol Chem. 1985 Aug 25;260(18):10346–10352. [PubMed] [Google Scholar]
  36. Silverstein R. L., Nachman R. L. Thrombospondin-plasminogen interactions: modulation of plasmin generation. Semin Thromb Hemost. 1987 Jul;13(3):335–342. doi: 10.1055/s-2007-1003509. [DOI] [PubMed] [Google Scholar]
  37. Strickland S., Reich E., Sherman M. I. Plasminogen activator in early embryogenesis: enzyme production by trophoblast and parietal endoderm. Cell. 1976 Oct;9(2):231–240. doi: 10.1016/0092-8674(76)90114-8. [DOI] [PubMed] [Google Scholar]
  38. Sutherland A. E., Calarco P. G., Damsky C. H. Expression and function of cell surface extracellular matrix receptors in mouse blastocyst attachment and outgrowth. J Cell Biol. 1988 Apr;106(4):1331–1348. doi: 10.1083/jcb.106.4.1331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Taraboletti G., Roberts D. D., Liotta L. A. Thrombospondin-induced tumor cell migration: haptotaxis and chemotaxis are mediated by different molecular domains. J Cell Biol. 1987 Nov;105(5):2409–2415. doi: 10.1083/jcb.105.5.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tuszynski G. P., Rothman V., Murphy A., Siegler K., Smith L., Smith S., Karczewski J., Knudsen K. A. Thrombospondin promotes cell-substratum adhesion. Science. 1987 Jun 19;236(4808):1570–1573. doi: 10.1126/science.2438772. [DOI] [PubMed] [Google Scholar]
  41. Varani J., Dixit V. M., Fligiel S. E., McKeever P. E., Carey T. E. Thrombospondin-induced attachment and spreading of human squamous carcinoma cells. Exp Cell Res. 1986 Dec;167(2):376–390. doi: 10.1016/0014-4827(86)90178-3. [DOI] [PubMed] [Google Scholar]
  42. Varani J., Nickoloff B. J., Riser B. L., Mitra R. S., O'Rourke K., Dixit V. M. Thrombospondin-induced adhesion of human keratinocytes. J Clin Invest. 1988 May;81(5):1537–1544. doi: 10.1172/JCI113486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Vestweber D., Gossler A., Boller K., Kemler R. Expression and distribution of cell adhesion molecule uvomorulin in mouse preimplantation embryos. Dev Biol. 1987 Dec;124(2):451–456. doi: 10.1016/0012-1606(87)90498-2. [DOI] [PubMed] [Google Scholar]
  44. Vischer P., Völker W., Schmidt A., Sinclair N. Association of thrombospondin of endothelial cells with other matrix proteins and cell attachment sites and migration tracks. Eur J Cell Biol. 1988 Oct;47(1):36–46. [PubMed] [Google Scholar]
  45. Wartiovaara J., Leivo I., Vaheri A. Expression of the cell surface-associated glycoprotein, fibronectin, in the early mouse embryo. Dev Biol. 1979 Mar;69(1):247–257. doi: 10.1016/0012-1606(79)90289-6. [DOI] [PubMed] [Google Scholar]
  46. Whitten W. K., Biggers J. D. Complete development in vitro of the pre-implantation stages of the mouse in a simple chemically defined medium. J Reprod Fertil. 1968 Nov;17(2):399–401. doi: 10.1530/jrf.0.0170399. [DOI] [PubMed] [Google Scholar]
  47. Wight T. N., Raugi G. J., Mumby S. M., Bornstein P. Light microscopic immunolocation of thrombospondin in human tissues. J Histochem Cytochem. 1985 Apr;33(4):295–302. doi: 10.1177/33.4.3884704. [DOI] [PubMed] [Google Scholar]
  48. Wilson I. B., Jenkinson E. J. Blastocyst differentiation in vitro. J Reprod Fertil. 1974 Jul;39(1):243–249. doi: 10.1530/jrf.0.0390243. [DOI] [PubMed] [Google Scholar]
  49. Wu T. C., Wan Y. J., Chung A. E., Damjanov I. Immunohistochemical localization of entactin and laminin in mouse embryos and fetuses. Dev Biol. 1983 Dec;100(2):496–505. doi: 10.1016/0012-1606(83)90242-7. [DOI] [PubMed] [Google Scholar]

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